Daniel:Notebook/HiResChrPaint/2013-5-11: Difference between revisions
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==DpnII Digestion== | ==DpnII Digestion== | ||
[[Daniel:Reagents/DpnII|Info]] | [[Daniel:Reagents/DpnII|Enzyme Info]] | ||
1. Prepared samples according to table | 1. Prepared samples according to table | ||
*DpnII-Lot 0091201; Buffer 0031105 | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt;font-weight:bold" align="center" | |||
| width="65" height="40" | Sample | |||
| width="65" | V4A | |||
| width="65" | V4B | |||
| width="65" | V4C | |||
| width="65" | V4D | |||
| width="65" | V6A | |||
| width="65" | V6B | |||
| width="65" | V6C | |||
| width="65" | V6D | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ng/uL | |||
| align="center" | 127.5 | |||
| align="center" | 156.3 | |||
| align="center" | 130.2 | |||
| align="center" | 130.6 | |||
| align="center" | 142.7 | |||
| align="center" | 138.9 | |||
| align="center" | 146.4 | |||
| align="center" | 133.4 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | ng | |||
| align="center" | 5100 | |||
| align="center" | 6252 | |||
| align="center" | 5208 | |||
| align="center" | 5224 | |||
| align="center" | 5708 | |||
| align="center" | 5556 | |||
| align="center" | 5856 | |||
| align="center" | 5336 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | uL | |||
| align="center" | 35 | |||
| align="center" | 35 | |||
| align="center" | 35 | |||
| align="center" | 35 | |||
| align="center" | 35 | |||
| align="center" | 35 | |||
| align="center" | 35 | |||
| align="center" | 35 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="35" | uL 10x buffer | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
| align="center" | 10 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="32" | uL DpnII (50U/uL) | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
| align="center" | 2 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | uL nf H20 | |||
| align="center" | 53 | |||
| align="center" | 53 | |||
| align="center" | 53 | |||
| align="center" | 53 | |||
| align="center" | 53 | |||
| align="center" | 53 | |||
| align="center" | 53 | |||
| align="center" | 53 | |||
|- style="font-size:12pt" align="center" | |||
|style="font-weight:bold" height="15" | Total | |||
| align="center" | 100 | |||
| align="center" | 100 | |||
| align="center" | 100 | |||
| align="center" | 100 | |||
| align="center" | 100 | |||
| align="center" | 100 | |||
| align="center" | 100 | |||
| align="center" | 100 | |||
|} | |||
2. Incubated 16 hours at 37C, followed by 20 minutes at 65C; Continued on [[Daniel:Notebook/HiResChrPaint/2013-5-12|5/12/2013]] |
Latest revision as of 21:34, 13 May 2013
Probe Prep Test 2 (Started 5/10/2013)[edit]
Ethanol Precipitation[edit]
7. Centrifuged 10000 rpm for 30 minutes at 4C 8. Removed supernatant added 750 uL 75% EtOH 9. Centrifuged 10000 rpm for 12 minutes at 4C 10. Removed supernatant, dried in hood 11. Resuspended with 50 uL 12. Column Purification, eluted with 50 uL 13. measured concentration in nanodrop
Nanodrop Results[edit]
Sample | V4A | V4B | V4C | V4D | V6A | V6B | V6C | V6D |
ng/uL | 127.5 | 156.3 | 130.2 | 130.6 | 142.7 | 138.9 | 146.4 | 133.4 |
ng | 5100 | 6252 | 5208 | 5224 | 5708 | 5556 | 5856 | 5336 |
ug in 4 sets | 21.8 | 22.5 |
DpnII Digestion[edit]
1. Prepared samples according to table *DpnII-Lot 0091201; Buffer 0031105
Sample | V4A | V4B | V4C | V4D | V6A | V6B | V6C | V6D |
ng/uL | 127.5 | 156.3 | 130.2 | 130.6 | 142.7 | 138.9 | 146.4 | 133.4 |
ng | 5100 | 6252 | 5208 | 5224 | 5708 | 5556 | 5856 | 5336 |
uL | 35 | 35 | 35 | 35 | 35 | 35 | 35 | 35 |
uL 10x buffer | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
uL DpnII (50U/uL) | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 |
uL nf H20 | 53 | 53 | 53 | 53 | 53 | 53 | 53 | 53 |
Total | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 |
2. Incubated 16 hours at 37C, followed by 20 minutes at 65C; Continued on 5/12/2013